Application of UL43 protein in preparing medicine for preventing and treating cellular mitochondrial dysfunction
A dysfunctional and mitochondrial technology, applied in the field of protein research and application mechanism, can solve the problems of inhibiting virus intercellular transmission, assembly, release, and affecting the endocytosis of viral envelope proteins, so as to increase the ATP content and broaden the application field.
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Embodiment 1
[0025] Example 1 HSV-1 UL43 protein endocytosis and intracellular localization experiments
[0026] UL43 protein is a multi-transmembrane protein. After HSV-1 successfully infects host cells, UL43 protein can be expressed on the host cell membrane. Preliminary studies have shown that the amino acid sequence of UL43 protein contains an endocytic motif. Therefore, this example mainly explores whether the HSV-1 UL43 protein depends on the YXXΦ motif for endocytosis from the host cell membrane into the cell, and the intracellular localization of the UL43 protein.
[0027] Experimental Materials:
[0028] HSV-1 UL43 eukaryotic expression plasmid pHUL43, HSV-1 UL43 (Y144A) mutant expression plasmid pHUL43 (Y144A), Flag-labeled UL43 expression plasmid pCMV(f)UL43, Flag-labeled UL43 mutant plasmid pCMV(f ) UL43 (Y144A). The eukaryotic expression vector pCDNA3.1+ was purchased from Invitrogen, and the plasmid pFLAG-CMV-2 was purchased from Sigma.
[0029] Herpes simplex virus F stra...
Embodiment 2
[0140] Example 2 Study on the Intracellular Function of HSV-1 UL43 Protein
[0141] It can be seen from Example 1 that the main organ of UL43 multi-transmembrane protein in the host cell is the mitochondria, and the mitochondria are the energy supply organs of the cells, providing a large amount of ATP for the life activities of the cells. Therefore, this part mainly explores the influence of HSV-1 UL43 protein on the mitochondrial production pathway of host cells, and the research on the localization target of UL43 on mitochondria.
[0142] Table 3 main reagents
[0143] Experimental Materials
source
Glucose detection and analysis kit
BioVision
ATP Content Assay Kit
Shanghai Kemin Biotechnology Co., Ltd.
Pyruvate Kinase Kit
Shanghai Baili Biotechnology Co., Ltd.
Fructose-6-Phosphate Kinase Activity Assay Kit
Shanghai Solaibao Biotechnology Co., Ltd.
Shanghai Baili Biotechnology Co., ...
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